CN116121221A - Bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof - Google Patents
Bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof Download PDFInfo
- Publication number
- CN116121221A CN116121221A CN202310054334.6A CN202310054334A CN116121221A CN 116121221 A CN116121221 A CN 116121221A CN 202310054334 A CN202310054334 A CN 202310054334A CN 116121221 A CN116121221 A CN 116121221A
- Authority
- CN
- China
- Prior art keywords
- antibacterial peptide
- dead
- bacillus cereus
- antibacterial
- rna helicase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000003910 polypeptide antibiotic agent Substances 0.000 title claims abstract description 103
- 241000193755 Bacillus cereus Species 0.000 title claims abstract description 31
- 102000007120 DEAD-box RNA Helicases Human genes 0.000 title claims abstract description 28
- 108010033333 DEAD-box RNA Helicases Proteins 0.000 title claims abstract description 28
- 241000607272 Vibrio parahaemolyticus Species 0.000 claims abstract description 47
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 30
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 18
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 14
- 239000003674 animal food additive Substances 0.000 claims abstract description 12
- 239000005452 food preservative Substances 0.000 claims abstract description 11
- 235000019249 food preservative Nutrition 0.000 claims abstract description 11
- 239000003814 drug Substances 0.000 claims abstract description 3
- 230000000844 anti-bacterial effect Effects 0.000 claims description 18
- 230000002147 killing effect Effects 0.000 claims description 12
- 239000003242 anti bacterial agent Substances 0.000 claims description 10
- 239000004480 active ingredient Substances 0.000 claims description 8
- 229940124350 antibacterial drug Drugs 0.000 claims description 5
- 239000004599 antimicrobial Substances 0.000 claims 1
- 238000009360 aquaculture Methods 0.000 claims 1
- 244000144974 aquaculture Species 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 10
- 230000005764 inhibitory process Effects 0.000 abstract description 4
- 208000015181 infectious disease Diseases 0.000 abstract description 3
- 229940079593 drug Drugs 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 20
- 210000000170 cell membrane Anatomy 0.000 description 15
- 241000894006 Bacteria Species 0.000 description 14
- 230000001580 bacterial effect Effects 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 11
- 108700042778 Antimicrobial Peptides Proteins 0.000 description 10
- 102000044503 Antimicrobial Peptides Human genes 0.000 description 10
- 108090000765 processed proteins & peptides Proteins 0.000 description 10
- 235000018102 proteins Nutrition 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 7
- 239000008363 phosphate buffer Substances 0.000 description 7
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 6
- 229940088710 antibiotic agent Drugs 0.000 description 6
- 235000013305 food Nutrition 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 102000004196 processed proteins & peptides Human genes 0.000 description 6
- 230000035699 permeability Effects 0.000 description 5
- 108020000946 Bacterial DNA Proteins 0.000 description 4
- 206010016952 Food poisoning Diseases 0.000 description 4
- 208000019331 Foodborne disease Diseases 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 229920001184 polypeptide Polymers 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 101100379209 Arabidopsis thaliana APD3 gene Proteins 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 244000000010 microbial pathogen Species 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 230000033077 cellular process Effects 0.000 description 2
- 238000001142 circular dichroism spectrum Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 239000000147 enterotoxin Substances 0.000 description 2
- 231100000655 enterotoxin Toxicity 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 230000000415 inactivating effect Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 108091065081 DEAD box helicase family Proteins 0.000 description 1
- 102000039002 DEAD box helicase family Human genes 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 208000005577 Gastroenteritis Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101001037191 Homo sapiens Hyaluronan synthase 1 Proteins 0.000 description 1
- 102100040203 Hyaluronan synthase 1 Human genes 0.000 description 1
- 108010028921 Lipopeptides Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 102000006833 Multifunctional Enzymes Human genes 0.000 description 1
- 108010047290 Multifunctional Enzymes Proteins 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 239000003570 air Substances 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000002390 cell membrane structure Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000013480 data collection Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 244000053095 fungal pathogen Species 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000002631 hypothermal effect Effects 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000024241 parasitism Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 1
- BJDYCCHRZIFCGN-UHFFFAOYSA-N pyridin-1-ium;iodide Chemical compound I.C1=CC=NC=C1 BJDYCCHRZIFCGN-UHFFFAOYSA-N 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000000304 virulence factor Substances 0.000 description 1
- 230000007923 virulence factor Effects 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3571—Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y306/00—Hydrolases acting on acid anhydrides (3.6)
- C12Y306/04—Hydrolases acting on acid anhydrides (3.6) acting on acid anhydrides; involved in cellular and subcellular movement (3.6.4)
- C12Y306/04013—RNA helicase (3.6.4.13)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Food Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Public Health (AREA)
- Biochemistry (AREA)
- Animal Husbandry (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- General Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Nutrition Science (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Insects & Arthropods (AREA)
- Marine Sciences & Fisheries (AREA)
- Birds (AREA)
- Peptides Or Proteins (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
The invention discloses a bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, the amino acid sequence of which is RKLLQFAKKLGIVFTK, and the molecular weight of the antibacterial peptide DB16 is 1890.337 daltons. The antibacterial peptide DB16 has obvious inhibition effect on vibrio parahaemolyticus and staphylococcus aureus, and can be used for preparing medicines, food preservatives or feed additives for preventing or inhibiting the infection of vibrio parahaemolyticus and staphylococcus aureus.
Description
Technical Field
The invention relates to the technical field of biology, in particular to bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof.
Background
In recent years, pathogenic microorganisms caused by bacterial contamination seriously affect human health and food safety, and food poisoning caused by food-borne pathogenic microorganisms has become one of the food safety problems.
Staphylococcus aureus is a common food-borne pathogenic bacterium and widely exists in the natural world such as air, dust, water resources and the like and in food processing environments. It can utilize the nutrients of protein, sugar, fat and vitamins in food to make propagation and metabolism, so that the nutritive value and quality of food are reduced, its quality-protecting period is shortened, and its quality is deteriorated. In addition, under proper conditions, staphylococcus aureus produces heat-resistant enterotoxins, and after the human body ingests foods contaminated by staphylococcus aureus or enterotoxins, symptoms of food poisoning such as nausea, vomiting, abdominal pain, shock, collapse, hypothermia and the like can be induced.
Vibrio parahaemolyticus is a swimming, non-sporulating, rod-shaped gram-negative bacterium of Proteus, has halophilic and mesophilic properties, and food poisoning event caused by the vibrio parahaemolyticus accounts for more than 60% of aquatic food poisoning. The bacterium invades host cells (intestinal epithelial cells) to release a large amount of virulence factors in vivo, so that the host cells die, and gastroenteritis and other diseases are caused.
Antibiotics are an important advancement in the medical history of humans for the treatment of bacterial diseases, but due to the abuse of antibiotics, bacterial resistance to antibiotics is becoming more and more serious, posing a serious threat to human health. The antibacterial peptide is also called host defensive peptide, is a small molecule polypeptide which is produced by organism under the induction of external stimulus, is coded and synthesized by specific genes and has broad-spectrum antibacterial activity or immunoregulatory function. Compared with antibiotics, the antibacterial peptide has the characteristics of broad spectrum, rapidness, high selectivity and low toxicity, and has multiple action targets, so that bacteria are difficult to generate drug resistance. Thus, antibacterial peptides are considered to be a good alternative to antibiotics.
Bacillus cereus (Bacillus cereus) is a gram-positive, rod-shaped, and spore-forming facultative anaerobic bacterium, often found in soil, water, and animal intestines, which can produce toxins that cause different types of gastrointestinal or other disease pathogens. The bacillus can also produce antagonism to fungal pathogens by antibacterial, nutrition competition, site exclusion, parasitism or induction methods, and can produce various active substances, such as low molecular polypeptides, lipopeptides antibiotics, antibacterial proteins and the like, which can effectively inhibit the growth of various pathogenic microorganisms, thus having good research and application values. The DEAD-box helicase family is a class of functional proteins found in host cells and is a broad class of ATP-dependent RNA helicases that, through binding to RNA and interaction, promote RNA folding and/or conformational rearrangement, play a critical role in a variety of cellular processes such as transcription, splicing, mRNA synthesis and translation. Members of this family possess the ability to recognize RNA and are involved in multiple cellular processes, so they can influence the natural immune response elicited upon infection of host cells by viruses in a variety of ways. Therefore, searching antibacterial peptide with antibacterial activity from DEAD-box helicase and exploring the antibacterial mechanism thereof is a technical problem to be solved in the field.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof, and solves the problems in the background art.
One of the technical schemes adopted for solving the technical problems is as follows: provides a bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, the amino acid sequence of which is shown in SEQ ID NO:1 is shown as follows:
RKLLQFAKKLGIVFTK
the molecular weight of the bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 was 1890.337 daltons, the positive charge was +5, the total hydrophobicity ratio was 50%, and the peptide was predicted to form an alpha helix by APD 3.
The second technical scheme adopted by the invention for solving the technical problems is as follows: provides an application of bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 in preparing antibacterial drugs for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
The third technical scheme adopted by the invention for solving the technical problems is as follows: the active ingredients of the antibacterial medicament comprise bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
in a preferred embodiment of the present invention, the active ingredient of the antibacterial agent is bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
in a preferred embodiment of the present invention, the antibacterial agent is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
The fourth technical scheme adopted for solving the technical problems is as follows: the application of the DEAD-box RNA helicase protein antibacterial peptide DB16 of bacillus cereus in preparing a food preservative for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus is provided.
The fifth technical scheme adopted by the invention for solving the technical problems is as follows: the effective components of the food preservative comprise bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, wherein the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
in a preferred embodiment of the present invention, the effective component of the food preservative is bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
in a preferred embodiment of the present invention, the food preservative is used for inhibiting and/or killing one or more of vibrio parahaemolyticus, staphylococcus aureus.
The sixth technical scheme adopted by the invention for solving the technical problems is as follows: provides an application of bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 in preparing an aquatic feed additive for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
The seventh technical scheme adopted by the invention for solving the technical problems is as follows: the effective components of the aquatic feed additive comprise bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, wherein the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
in a preferred embodiment of the present invention, the effective component of the aquatic feed additive is bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
in a preferred embodiment of the invention, the aquatic feed additive is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
The invention uses APD3, pymol2.0, SWISS-MODEL and other software and websites to carry out bioinformatics prediction by taking the protein sequence of DEAD-box RNA helicase protein related to immunity in bacillus cereus as an object, thus obtaining a polypeptide DB16 with a novel amino acid sequence. Studying the antibacterial activity of the polypeptide DB16 on vibrio parahaemolyticus and staphylococcus aureus; taking vibrio parahaemolyticus as an example, observing the damage degree of DB16 to bacterial cell membranes by using flow cytometry and PI staining; meanwhile, the genome DNA of vibrio parahaemolyticus is extracted, and the influence of the genome DNA on bacterial DNA is verified. Experimental results show that the peptide has obvious inhibition effect on vibrio parahaemolyticus and staphylococcus aureus.
The antibacterial mechanism is to act on the cell membrane to deform and shrink the cell membrane, enhance the permeability of the cell membrane to enable the peptide to pass through the cell membrane of the bacteria, combine with the DNA of the bacteria, inhibit the replication and synthesis of the DNA and the like, thereby achieving the effect of inactivating the bacteria.
The antimicrobial peptides of the invention can be synthesized using methods known to those skilled in the art, such as solid phase synthesis, and purified using methods known to those skilled in the art, such as high performance liquid chromatography.
The implementation of the invention has the following beneficial effects:
the antibacterial peptide DB16 has obvious inhibition effect on vibrio parahaemolyticus and staphylococcus aureus. The antibacterial peptide DB16 can be prepared into antibacterial drugs, food preservatives or aquatic feed additives for preventing or treating diseases caused by infection of vibrio parahaemolyticus and staphylococcus aureus.
Drawings
FIG. 1 is a schematic structural diagram of an antibacterial peptide DB16.
FIG. 2 is a graph showing a control of the measurement of the Minimum Bactericidal Concentration (MBC) of the antibacterial peptide DB16 against Vibrio parahaemolyticus. Wherein the concentrations of the antimicrobial peptide DB16 in the graphs A-I are 0 mug/mL, 500 mug/mL, 250 mug/mL, 125 mug/mL, 62.5 mug/mL, 31.25 mug/mL, 15.625 mug/mL, 7.813 mug/mL and 3.9 mug/mL, respectively.
FIG. 3 is a graph showing a control of the Minimum Bactericidal Concentration (MBC) of the antibacterial peptide DB16 against Staphylococcus aureus. Wherein the concentrations of the antimicrobial peptide DB16 in the graphs A-I are 0 mug/mL, 500 mug/mL, 250 mug/mL, 125 mug/mL, 62.5 mug/mL, 31.25 mug/mL, 15.625 mug/mL, 7.813 mug/mL and 3.9 mug/mL, respectively.
FIG. 4 is a time-kill kinetics plot of the antimicrobial peptide DB16 against Vibrio parahaemolyticus.
Fig. 5 is a time kill kinetics plot of the antimicrobial peptide DB16 against staphylococcus aureus.
FIG. 6 is a graph showing the effect of the antibacterial peptide DB16 on the permeability of cell membranes of Vibrio parahaemolyticus (protein leakage).
FIG. 7 is a graph showing the effect of the antibacterial peptide DB16 on the permeability of cell membranes of Vibrio parahaemolyticus.
FIG. 8 is a graph showing the circular dichroism spectrum of the antimicrobial peptide DB16 under different conditions.
FIG. 9 is a gel electrophoresis chart of the antibacterial peptide DB16 and Vibrio parahaemolyticus genomic DNA.
Wherein bands 1-10: the mass ratio of the antibacterial peptide DB16 to the DNA is 25/4, 25/6, 25/10, 25/12, 25/14, 25/16, 25/18, 25/20, 25/22, 25/25 and the band 11 are the DNA of a control group.
FIG. 10 is a graph showing a circular dichroism spectrum after binding of the antibacterial peptide DB16 to Vibrio parahaemolyticus genomic DNA.
Detailed Description
For a better understanding of the present invention, reference will now be made in detail to the following examples and accompanying drawings, which are included to provide a further understanding of the invention, and it is to be understood by those skilled in the art that the following examples are not intended to limit the scope of the invention.
The experimental methods used in the following examples are conventional methods unless otherwise specified.
Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
EXAMPLE 1 screening of antibacterial peptide DB16
The bioinformatics prediction is carried out by using software such as APD3, pymol2.0 and SWISS-MODEL, and websites by taking the protein sequence of DEAD-box RNA helicase protein related to immunity in bacillus cereus as an object, the charge and hydrophobicity of the amino acid sequence are analyzed by using APD3 (table 1), the three-dimensional structure is predicted by using Pymol2.0 and Swiss-MODEL (shown in figure 1), and finally the amino acid sequence RKLLQFAKKLGIVFTK with the antibacterial performance is screened out and chemically synthesized (synthesized by Beijing midbody matte biological technology Co., ltd.) and the antibacterial activity is verified.
Table 1: predictive analysis of antibacterial peptides in bacillus cereus
Example 2 determination of Minimum Bactericidal Concentration (MBC) of antibacterial peptide DB16
Culturing Vibrio parahaemolyticus and Staphylococcus aureus (Vibrio parahaemolyticus ATCC17802 and Staphylococcus aureus ATCC27217 are all laboratory-preserved strains) at 37deg.C for 12 hr to logarithmic phase, and diluting to 10 in 0.01M pH 7.2 phosphate buffer 5-6 CFU/mL. Antibacterial peptide DB16 was dissolved in phosphate buffer, mixed with bacteria in equal volume at 37℃for 2h incubation. The minimum inhibitory concentration (MBC) refers to the minimum concentration of antimicrobial peptide that kills bacteria after incubation at 37 ℃. As shown in FIGS. 2 and 3, the Minimum Bactericidal Concentration (MBC) of the antibacterial peptide DB16 against Vibrio parahaemolyticus and Staphylococcus aureus was 7.8125. Mu.g/mL and 3.90625. Mu.g/mL, respectively.
Example 3 Time-sterilization Curve (Time kill) determination of antibacterial peptide DB16
Culturing Vibrio parahaemolyticus and Staphylococcus aureus at 37deg.C for 12 hr to logarithmic phase, and diluting to 10 in 0.01M pH 7.2 phosphate buffer 5-6 CFU/mL. Antibacterial peptide DB16 with 1 XMBC concentration was taken and incubated at 37 ℃ in equal volume with bacteria, each plate was sampled every 30 minutes, and the total number of colonies was recorded after incubation at 37 ℃ overnight. As shown in fig. 4 and 5, the antibacterial peptide DB16 can kill vibrio parahaemolyticus within 1.5 hours and staphylococcus aureus within 0.5 hour.
Example 4 Effect of antibacterial peptide DB16 on Vibrio parahaemolyticus cell membrane permeability
Protein leakage: 200 mu L of the vibrio parahaemolyticus strain preserved at-20 ℃ is inoculated into LB culture medium, and shake culture is carried out for 10 hours at 37 ℃ and 200r/h until the logarithmic growth phase of the bacteria. After preparing a bacterial suspension of 0.01M phosphate buffer, mixing with antibacterial peptide DB16 with concentration of 1 XMBC and 2 XMBC in equal proportion, and measuring OD of the bacterial suspension at intervals of 0-2h every 10min by using a multifunctional enzyme-labeled instrument 280nm The protein leakage amount of the cells was examined. The experiment was run with equal amounts of 0.01M phosphate buffer mixed with bacteria in equal proportions as a blank (fig. 6).
Detection of absorbance at 280nm can be used to estimate the amount of protein leaking from the cytoplasm. As shown in FIG. 6, the amount of protein in the culture solution treated with the antibacterial peptide DB16 increased in a dose-dependent manner as the concentration of the antibacterial peptide DB16 increased.
Flow cytometry: vibrio parahaemolyticus is cultured at 37 ℃ for 12h to logarithmic growth phase, 1mL of bacterial liquid is taken and centrifuged for 1min at 12000r/min in a 1.5mL centrifuge tube, supernatant is removed, and the suspension is resuspended to 1mL of sterile 0.01M phosphate buffer solution and repeated for 3 times. After preparing a bacterial suspension of 0.01M phosphate buffer, antibacterial peptide DB16 with concentration of 1 XMBC and 2 XMBC was mixed with bacterial solution in equal proportion (more than 200. Mu.L), and incubated in a biochemical incubator at 37℃for 2 hours, using 0.01M phosphate buffer as a blank control group. Adding PI dye with an equal volume concentration of 50 mug/mL into the mixed solution after incubation is completed by using Pyridine Iodide (PI) as fluorescent dye, incubating for 15min at 4 ℃, and loading to flowAnd detecting by a cytometer. Detecting each tube is set to obtain at least 10 4 Bacterial cells. A data collection analysis protocol was set up and samples were analyzed by detecting scattered light Signals (SC) and propidium iodide fluorescent signals (PI) (fig. 7).
As shown in fig. 7, the control group has a vibrio parahaemolyticus mortality rate of only 11.7%, which indicates that the cell membrane structure is complete, PI cannot penetrate the cell membrane to enter the cell interior, and the bacterial mortality rate is low; the death rate of the vibrio parahaemolyticus added with the antibacterial peptide DB16 with the concentration of 1 XMBC is obviously increased to 94.1 percent; vibrio parahaemolyticus mortality in the experimental group to which 2 XMBC concentration of antibacterial peptide was added was slightly increased up to 98.9% compared to 1 XMBC. It can be seen that the concentration of the antibacterial peptide DB16 positively correlated with the mortality of Vibrio parahaemolyticus and increased in a dose-dependent manner.
EXAMPLE 5 round dichroism determination of antibacterial peptide DB16 secondary Structure
The average residue molar ellipticity of the antibacterial peptide DB16 was determined with a Jasco810 spectropolarimeter (Jasco, tokyo) CD at 25℃at a scan rate of 100 nm/min. Antibacterial peptide DB16 was dissolved in 0.01M PBS and 25mM Sodium Dodecyl Sulfate (SDS) to a final concentration of 0.20mg/mL, and the spectra were scanned from 180 to 280nm with two scans.
As shown in fig. 8, in PBS environment, the antimicrobial peptide DB16 appeared to have a negative peak at 198nm and a negative peak at 220nm, indicating that the secondary conformation of the antimicrobial peptide DB16 was predominantly random coil in PBS environment. In SDS environment, there is positive peak at 193nm, and two negative peaks at 218nm and 220nm, which indicates that the secondary conformation is mainly alpha-helix in SDS environment, and the antibacterial peptide may have structural transformation when contacting with bacterial cell membrane.
Example 6 interaction of antibacterial peptide DB16 with bacterial DNA
The interaction of the antibacterial peptide DB16 with Vibrio parahaemolyticus genomic DNA was studied using a DNA gel blocking method. Vibrio parahaemolyticus was cultured in 50mL of nutrient broth at 37℃for 12h, and the ratio of Optical Density (OD) of bacteria 260 and 280nm was used 260 /OD 280 1.90) the purity of the extracted genomic DNA was evaluated. Next, 10. Mu.L of DNA (20 ng/. Mu.L) was mixed with the antibacterial peptide DB16 at 25 ℃Mixing, namely, the mass ratio of the antibacterial peptide DB16 to the DNA is respectively 25/4, 25/6, 25/10, 25/12, 25/14, 25/16, 25/18, 25/20, 25/22, 25/25 and 0, and incubating at 37 ℃ for 1.5 hours after uniformly mixing. Then 8 μl each was electrophoresed on a 1% agarose gel and gel blocking was observed under ultraviolet irradiation using a GelDoc XR gel imaging system (Bio-Rad, usa).
As shown in FIG. 9, the DNA bands did not run out at the ratios of 25/4, 25/6, 25/8, 25/12, and the band clarity and brightness increased significantly with increasing ratios of 25/14, 25/16, 25/18, 25/20, 25/22, 25/25, respectively; gel electrophoresis of the control Vibrio parahaemolyticus genomic DNA is shown as the brightest and clear band. The antibacterial peptide BaD/DNA mass ratio is shown as follows: 25/4, 25/6, 25/8, 25/12, the DNA is blocked and destroyed, the band does not run out, but the band gradually and clearly runs out as the concentration of the antimicrobial peptide decreases.
Example 7 Effect of antibacterial peptide DB16 on its secondary Structure by action with bacterial DNA
The average residue molar ellipticity of the antibacterial peptide DB16 was determined again using a Jasco810 spectropolarimeter (Jasco, tokyo) CD at 25℃at a scan rate of 100 nm/min. Antibacterial peptide DB16 was dissolved in 25mM Sodium Dodecyl Sulfate (SDS) to a final concentration of 0.20mg/mL, and Vibrio parahaemolyticus DNA (20 ng/. Mu.L) was mixed with antibacterial peptide DB16 at 25℃for incubation for 2 hours, and then the spectra thereof were scanned from 180 to 280nm by two scans (FIG. 10).
After incubation of DNA with the antibacterial peptide DB16, positive peaks at 193nm and negative peaks at 218nm and 220nm still exist, but the spectrogram of the antibacterial peptide DB16 is expanded as a whole, which shows that the antibacterial peptide DB16 has an interaction relationship with the vibrio parahaemolyticus genomic DNA.
In conclusion, the invention provides a brand-new antibacterial peptide DB16, and the Minimum Bactericidal Concentration (MBC) of the antibacterial peptide DB16 on vibrio parahaemolyticus and staphylococcus aureus is 7.8125 mug/mL and 3.90625 mug/mL respectively, which shows that the antibacterial peptide DB16 has a strong inhibition effect on vibrio parahaemolyticus and staphylococcus aureus. The research shows that the antibacterial peptide DB16 acts on cell membranes to deform and shrink the cell membranes, enhances the permeability of the cell membranes to enable the cell membranes to penetrate through bacterial cell membranes, combines with bacterial DNA, inhibits the functions of DNA replication and synthesis and the like, thereby achieving the effect of inactivating bacteria.
While specific embodiments of the invention have been described above, it will be appreciated by those skilled in the art that the specific embodiments described are illustrative only and not intended to limit the scope of the invention, and that equivalent modifications and variations of the invention in light of the spirit of the invention will be covered by the claims of the present invention.
Claims (13)
1. A DEAD-box RNA helicase protein antibacterial peptide DB16 of bacillus cereus has an amino acid sequence shown in SEQ ID NO:1.
2. The use of bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 as claimed in claim 1 for the preparation of an antibacterial medicament, characterized in that: the antibacterial drug is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
3. An antibacterial agent characterized in that: the active ingredient of the antibacterial peptide is bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
4. an antimicrobial agent as claimed in claim 3 wherein: the active ingredient of the antibacterial peptide is bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
5. the antibacterial agent according to any one of claims 3 or 4, wherein: the antibacterial drug is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
6. The use of bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 as claimed in claim 1 for the preparation of a food preservative, characterized in that: the food preservative is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
7. A food preservative characterized by: the active ingredients of the antibacterial peptide comprise bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
8. the food preservative according to claim 7, characterized in that: the active ingredient of the antibacterial peptide is bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
9. the food preservative according to any one of claims 7 or 8, characterized in that: the antibacterial drug is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
10. Use of the bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 according to claim 1 for the preparation of an aquatic feed additive, characterized in that: the aquatic feed additive is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
11. An aquatic feed additive, characterized in that: the active ingredients of the antibacterial peptide comprise bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
12. an aquatic feed additive as claimed in claim 11 wherein: the active ingredient of the antibacterial peptide is bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16, and the amino acid sequence of the antibacterial peptide DB16 is SEQ ID NO:1.
13. an aquaculture feed additive according to any one of claims 11 or 12 wherein: the aquatic feed additive is used for inhibiting and/or killing one or more of vibrio parahaemolyticus and staphylococcus aureus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310054334.6A CN116121221B (en) | 2023-02-03 | Bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310054334.6A CN116121221B (en) | 2023-02-03 | Bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116121221A true CN116121221A (en) | 2023-05-16 |
| CN116121221B CN116121221B (en) | 2024-07-02 |
Family
ID=
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200048425A (en) * | 2018-10-30 | 2020-05-08 | 대한민국(관리부서:국립수산과학원) | Antimicrobial peptide derived from the phospholipase of rock bream and uses thereof |
| CN112898386A (en) * | 2021-03-02 | 2021-06-04 | 集美大学 | Large yellow croaker myosin heavy chain antibacterial peptide LCMHC and application thereof |
| KR20220037035A (en) * | 2020-09-16 | 2022-03-24 | 제주대학교 산학협력단 | Antibacterial peptide derived from Haliotis madaka |
| CN114957395A (en) * | 2022-01-05 | 2022-08-30 | 江西科诺生物科技有限公司 | Gene of bee antibacterial peptide and application thereof |
| WO2023274024A1 (en) * | 2021-06-28 | 2023-01-05 | 厦门大学 | Antibacterial peptide scyampcin44-63 and application thereof |
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200048425A (en) * | 2018-10-30 | 2020-05-08 | 대한민국(관리부서:국립수산과학원) | Antimicrobial peptide derived from the phospholipase of rock bream and uses thereof |
| KR20220037035A (en) * | 2020-09-16 | 2022-03-24 | 제주대학교 산학협력단 | Antibacterial peptide derived from Haliotis madaka |
| CN112898386A (en) * | 2021-03-02 | 2021-06-04 | 集美大学 | Large yellow croaker myosin heavy chain antibacterial peptide LCMHC and application thereof |
| WO2023274024A1 (en) * | 2021-06-28 | 2023-01-05 | 厦门大学 | Antibacterial peptide scyampcin44-63 and application thereof |
| CN114957395A (en) * | 2022-01-05 | 2022-08-30 | 江西科诺生物科技有限公司 | Gene of bee antibacterial peptide and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| MA DENGYING等: "Identification, expression and activity analyses of five novel duck beta-defensins", PLOS ONE, vol. 07, no. 10, 24 October 2012 (2012-10-24), pages 47743 * |
| ZHENG, Z: "ATP-dependent helicase, partial [Bacillus cereus]", 《GENBANK》, 登录号 MDA1570333.1, 10 January 2023 (2023-01-10) * |
| 宁亚维等: "抗菌肽brevilaterin与ε-聚赖氨酸对金黄色葡萄球菌的协同抑菌机理", 食品科学, vol. 41, no. 05, 12 December 2019 (2019-12-12), pages 15 - 22 * |
| 陈伟等: "抗菌肽的研究进展及在食品中的应用", 食品科技, vol. 43, no. 04, 20 April 2018 (2018-04-20), pages 65 - 69 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102071162B (en) | Bacillus subtilis LFB112 as well as bacteriocin produced by same and application thereof | |
| D'Alvise et al. | Inactivation of Vibrio anguillarum by attached and planktonic Roseobacter cells | |
| CN105175518B (en) | The bacteriocin and preparation method thereof that bacillus coagulans FM603 is generated | |
| CN114134083B (en) | Bacillus bailii and application thereof | |
| CN104945484B (en) | A kind of antibacterial peptide and preparation method and application | |
| CN112898386A (en) | Large yellow croaker myosin heavy chain antibacterial peptide LCMHC and application thereof | |
| CN114085789A (en) | Pediococcus pentosaceus MA.WTPQJ01 and application thereof | |
| CN107252475B (en) | The application of natural host defense peptide Alligatorin4 | |
| CN115536737A (en) | Application of cobra antibacterial peptide OH-CATH30 in resisting aquatic animal pathogenic bacteria | |
| CN113321708B (en) | Preparation of artificially designed antibacterial peptide and application of artificially designed antibacterial peptide in aquatic products | |
| CN102978126B (en) | Novel lactic acid bacterial strain, composition containing same and use thereof | |
| CN106883288B (en) | Antibacterial peptide BV21 and application thereof | |
| CN106632606B (en) | Antibacterial lipopeptide bacaucin derivative and application thereof in inhibiting bacterial infection | |
| CN116731153B (en) | Pig beta-interferon antibacterial peptide IFN7 and application thereof | |
| CN106674331B (en) | Antibacterial lipopeptide bacaucin and preparation method and application thereof | |
| CN116121221B (en) | Bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof | |
| CN116121221A (en) | Bacillus cereus DEAD-box RNA helicase protein antibacterial peptide DB16 and application thereof | |
| CN113087771B (en) | Penaeus vannamei DNA (deoxyribonucleic acid) combined antibacterial peptide VPDB40 and application thereof | |
| CN116640186A (en) | Human nasal fluid complement factor antibacterial peptide C4-AII and application thereof | |
| CN116622670A (en) | Bacillus cereus ATP synthase beta subunit antibacterial peptide BaD and application thereof | |
| CN106883289B (en) | Antibacterial peptide BV21 | |
| Lueders et al. | Unusual outbreak of fatal clostridiosis in a group of captive brown pelicans (Pelecanus occidentalis) | |
| Bocamdé et al. | Improvement of the growth performance, innate immunity and disease resistance of nile tilapia (Oreochromis niloticus) against vibrio parahaemolyticus 1T1 following dietary application of the probiotic strain Lactobacillus plantarum 1KMT | |
| CN112940998B (en) | Bacterial strain for preventing bacterial diseases and application | |
| CN117700486B (en) | Brevibacterium acetylicum fermentation liquor antibacterial peptide YX-2 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant |